|
|
- 2016
人CAP1蛋白与PIF1解螺旋酶及其剪接体蛋白的相互作用
|
Abstract:
摘要：目的 在细胞水平及体外验证腺苷酸环化酶相关蛋白1(CAP1)蛋白与PIF1解螺旋酶及其剪接体是否存在相互作用。方法 采用激光共聚焦免疫荧光实验，通过细胞内蛋白共定位确定蛋白质间的相互作用关系；GST Pull-down实验在体外验证CAP1与PIF1及其剪接体是否存在直接的相互作用。结果 激光共聚焦免疫荧光实验显示，CAP1与PIF1全长蛋白及其剪接体BC018978没有共定位，但与剪接体AF108138C存在共定位。GST Pull-down未检测到CAP1与PIF1全长及其剪接体的相互作用。结论 CAP1与PIF1剪接体蛋白可能存在非直接相互作用。
ABSTRACT: Objective?? To determine whether human CAP1 interacts with PIF1 helicase and its splice variants proteins in vivo and in vitro. Methods? ?The co-localization of protein was directly observed to determine the interaction between the proteins by confocal immunofluorescence assay. The GST Pull-down experiment was used to test the existence of a direct interaction between CAP1 protein and PIF1 or its splice variants proteins. Results?? By using confocal immunofluorescence assay, CAP1 protein and PIF1 protein or its splice variants BC018978 had no co-localization, but CAP1 and splice variants AF108138C were co-located in the cytoplasm. Experimental results of the GST Pull-down showed that there was not direct interaction between CAP1 and PIF1 or its splice variants proteins. Conclusion?? These results show that there may be indirect interaction between CAP1 and PIF1 splice variants proteins
| [1] | HANADA K, HICKSON ID. Molecular genetics of RecQ helicase disorders[J]. Cell Mol Life Sci, 2007, 64(17):2306-2322. |
| [2] | SHIMADA K, GASSER SM. DNA replication: PIF1 pulls the plug on stalled replication forks[J]. Curr Biol, 2012, 22(10):404-405. |
| [3] | WANG J, ENGLUND PT, JENSEN RE. TbPIF8, a trypanosoma brucei protein related to the yeast Pif1 helicase, is essential for cell viability and mitochondrial genome maintenance[J]. Molmicrobiol, 2012, 83(3):471-485. |
| [4] | TAN M, SONG XL, ZHANG GL, et al. Overexpression of adenylate cyclase-associated protein 1 is associated with metastasis of lung cancer[J]. Oncol Reports, 2013, 30(4):1639-1644. |
| [5] | HUDRY B, VIALA S, GRABA Y, et al. Visualization of protein interactions in living Drosophila embryos by the bimolecular fluorescence complementation assay[J]. BMC Biol, 2011, 9(1):1-18. |
| [6] | WALSH BW, LENHART JS, SCHROEDER JW, et al. Far Western blotting as a rapid and efficient method for detecting interactions between DNA replication and DNA repair proteins[J]. Methods Mol Biol, 2012, 922:161-168. |
| [7] | SATO Y, KAMEYA M, ARAI H, et al. Detecting weak proteinprotein interactions by modified far-Western blotting[J]. J Biosci Bioeng, 2011, 112(3):304-307. |
| [8] | BOCHMAN ML, SABOURI N, ZAKIAN VA. Unwinding the functions of the PIF1 family helicases[J]. DNA Repair (Amst), 2010, 9(3):237-249. |
| [9] | CHISHOLM KM, AUBERT SD, FREESE KP, et al. A genomewide screen for suppressors of Alu-mediated rearrangements reveals a role for PIF1[J]. PLoS One, 2012, 7(2):e30748. |
| [10] | HUBBERSTEY AV, MOTTILLO EP. Cyclase-associated proteins: CAPacity for linking signal transduction and actin polymerization[J]. FASEB J, 2002, 16(6):487-499. |
| [11] | ZUCCHI I, MENTO E, KUZNETSOV VA, et al. Gene expression profiles of epithelial cells microscopically isolated from a breast-invasive ductal carcinoma and a nodal metastasis[J]. Proc Nat Acad Sci United States Am, 2004, 101(52):18147-18152. |
| [12] | YAMAZAKI K, TAKAMURA M, MASUGI Y, et al. Adenylate cyclase-associated protein 1 overexpressed in pancreatic cancers is involved in cancer cell motility[J]. Lab Invest, 2009, 89(4):425-432. |
| [13] | BERENDZEN KW, BOHMER M, WALLMEROTH N, et al. Screening for in planta protein-protein interactions combining bimolecular fluorescence complementation with flow cytometry[J]. Plant Methods, 2012, 8(1):32-54. |
| [14] | SCHULDT A. DNA replication: Pif1 overcomes a quadruplex hurdle[J]. Nat Rev Mol Cell Biol, 2011, 12(7):402-403. |
| [15] | GAGOU ME, GANESH A, THOMPSON R, et al. Suppression of apoptosis by PIF1 helicase in human tumor cells[J]. Cancer Res, 2011, 71(14):4998-5008. |
| [16] | GEORGE T, WEN Q, GRIFFITHS R, et al. Human Pif1 helicase unwinds synthetic DNA structures resembling stalled DNA replication forks[J]. Nucleic Acids Res, 2009, 37(19):6491-6502. |
| [17] | SANDERS CM. Human Pif1 helicase is a G-quadruplex DNA-binding protein with G-quadruplex DNA-unwinding activity[J]. Biochem J, 2010, 430(1):119-128. |
| [18] | GU YQ, MASUDA Y, KAMIYA K. Biochemical analysis of human PIF1 helicase and functions of its N-terminal domain[J]. Nucleic Acid Res, 2008, 36(19):6296-6308. |
| [19] | VEZINA CM, WALKER NJ, OLSON JR. Subchronic exposure to TCDD, PeCDF, PCB126, and PCB153: effect on hepatic gene expression[J]. Environ Health Persp, 2004, 112(16):1636-1644. |
| [20] | SAINI N, RAMAKRISHNAN S, ELANGO R, et al. Migrating bubble during break-induced replication drives conservative DNA synthesis[J]. Nature, 2013, 502(7471):388-392. |
| [21] | GAGOU ME, GANESH A, PHEAR G, et al. Human PIF1 helicase supports DNA replication and cell growth under oncogenic-stress[J]. Oncotarget, 2014, 5(22):11381-11398. |
